Rab5 c8b1

Manufactured by Cell Signaling Technology

Rab5 (C8B1) is a primary antibody that recognizes the Rab5 protein. Rab5 is a small GTPase that plays a key role in the regulation of early endocytic pathways.

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3 protocols using rab5 c8b1

Characterization of Neisseria gonorrhoeae antibodies

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The purified chicken IgY anti-Neisseria gonorrhoeae antibody was raised against formalin-killed FA1090 gonococci by Cocalico Biologicals and it cross-reacts with the MS-11, FA19 and F62 strains. The rabbit anti-N.g polyclonal antibody (cat# 20-NR08) was purchased from Fitzgerald Industries. Rabbit monoclonal antibodies against the following antigens were purchased from Cell Signaling—Rab5 (C8B1), Rab7 (D95F2), EEA1 (C45B10), GM130 (D6B1), Ezrin (#3145), LC3A/B (D3U4C), V5-Tag (E9H80), CEACAM1 (D3R80) and ACTR2 (#3128S). Mouse antibodies against the following antigens were purchased from Santa Cruz–Actin (sc8432), FMNL1 (sc390466), FMNL2 (sc390298), FHOD1 (sc365473), DAAM1 (sc100942). Mouse anti-Cas9 (7A9-3A3) was purchased from Cell Signaling. Anti-FMNL3 (ab57963) and anti-Galectin 3 (ab2785) mouse monoclonal antibodies were purchased from Abcam. Anti-human LAMP-2 (H4B4) was purchased from BioLegend. Anti-DIAPH2 (A300-079A-T) was purchased from Bethyl Labs. Highly cross-absorbed Alexa fluorophore conjugated secondary antibodies were purchased from Life Technologies.
Inhibitors used in this study were purchased from: (1) Cayman Chemicals—cytochalasin D (5μM), CK869 (10μM), Dynasore (30μM), GSK269962 (1μM), Nocodazole (3μM), ML-141 (5μM), EHT 1864 (5μM), BMS-5 (1μM) and (-) blebbistatin (5μM); (2) Cell Signaling—LY294002 (10μM); (3) Abcam—SMIFH2 (12.5μM to 100μM).

Ivanov S.S., Castore R., Juarez Rodriguez M.D., Circu M, & Dragoi A.M. (2021). Neisseria gonorrhoeae subverts formin-dependent actin polymerization to colonize human macrophages. PLoS Pathogens, 17(12), e1010184.

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Vaccinia Virus Internalization Kinetics

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Cells were transfected with plasmids on coverslips in 24‐well plates and infected with VACV mCherry‐A4 (multiplicity of infection, MOI = 20) in FCS‐free medium. Virus was bound to cells for 30 min in a cold water bath. Cells were then washed and incubated in a 37°C water bath for indicated time points (up to 4 h). Bound non‐internalized particles were stained with the MAb 7D11 on ice without permeabilization. Fixed cells were analyzed with a Zeiss LSM 510 Meta confocal microscope using a 63× objective and seven to ten Z‐stacks were acquired per image. All images represent maximum projects when not indicated otherwise. Cells analyzed for endogenous markers were first subjected to 7D11 staining as above, fixed and permeabilized for 2 min with 0.1% Triton‐X‐100 prior to staining with antibodies directed against each marker, followed by Alexa Fluor 488 secondary antibody. Antibodies used were Rab5 [Cell Signaling; Rab5 (C8B1)], Rab7 [Cell Signaling; Rab7 (D95F2)], LAMP1 [(Abcam; LAMP1 (ab24170)] and EEA1 [Cell Signaling; EEA1 (3288)].

Rizopoulos Z., Balistreri G., Kilcher S., Martin C.K., Syedbasha M., Helenius A, & Mercer J. (2015). Vaccinia Virus Infection Requires Maturation of Macropinosomes. Traffic (Copenhagen, Denmark), 16(8), 814-831.

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Antibodies and Inhibitors for Gonococcal Trafficking

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The purified chicken IgY anti-Neisseria gonorrhoeae antibody was raised against formalin-killed FA1090 gonococci by Cocalico Biologicals and it cross-reacts with the MS-11, FA19 and F62 strains. The rabbit anti-N.g polyclonal antibody (cat# 20-NR08) was purchased from Fitzgerald Industries. Rabbit monoclonal antibodies against the following antigens were purchased from Cell Signaling -Rab5 (C8B1), Rab7 (D95F2), EEA1 (C45B10), GM130 (D6B1), Ezrin (#3145), LC3A/B (D3U4C), V5-Tag (E9H80), CEACAM1 (D3R80) and ACTR2 (#3128S).
Mouse antibodies against the following antigens were purchased from Santa Cruz -Actin (sc8432), FMNL1 (sc390466), FMNL2 (sc390298), FHOD1 (sc365473), DAAM1 (sc100942).
Mouse anti-Cas9 (7A9-3A3) was purchased from Cell Signaling. Anti-FMNL3 (ab57963) and anti-Galectin 3 (ab2785) mouse monoclonal antibodies were purchased from Abcam. Anti-human LAMP-2 (H4B4) was purchased from BioLegend. Anti-DIAPH2 (A300-079A-T) was purchased from Bethyl Labs. Highly cross-absorbed Alexa fluorophore conjugated secondary antibodies were purchased from Life Technologies.
Inhibitors used in this study were purchased from: (1) Cayman Chemicals -cytochalasin D (5µM), CK869 (10µM), Dynasore (30µM), GSK269962 (1µM), Nocodazole (3µM), ML-141 (5µM), EHT 1864 (5µM), BMS-5 (1µM) and (-) blebbistatin (5µM); (2) Cell Signaling -LY294002 (10µM); (3) Abcam -SMIFH2 (25µM).

Ivanov S.S., Castore R., Circu M.L., & Dragoi A. (2021). Neisseria gonorrhoeae subverts formin-dependent actin polymerization to colonize human macrophages.

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